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[Prokaryotic expression and characterization of the Rv2994 gene from Mycobacterium tuberculosis]
Ming-cai Zhao1, Lang Bao, Yue-han Wu
1Infection and Immunity Unit, West China Center of Medical Sciences, Sichuan University, Chengdu 610041, China.
Objective:
To construct a prokaryotic expression vector bearing Rv2994 gene from Mycobacterium tuberculosis and provide materials for investigating the function of the gene.
Methods:
The Rv2994 gene was amplified by Polymerase Chain Reaction from Mycobacterium tuberculosis H37Rv strain and cloned into prokaryotic expression vector pGEX-1lamdaXT. The recombinant plasmid pGEX-2994 was sequenced and transformed into E. coli JM109 to be induced with IPTG and expressed the 73kDa fusion protein GST-Rv2994. It's antigenicity was confirmed by Western blotting. The expression product was purified and immunized the new Zealand rabbits.
Results:
The Rv2994 gene was amplified accurately from the genome DNA of H37Rv. A recombinant fused expression vector pGEX-Rv2994 was constructed and GST-Rv2994 protein was purifiel to immunize New Zealand rabbit.
Conclusion:
The prokaryotic expression vector pGEX-2994 was constructed, and the 73kDa fusion protein GST-Rv2994 was expressed and purified successfully. It provided the basis for the further study of the Rv2994 gene.
